Anthrax toxin-mediated delivery of a cytotoxic T-cell epitope in vivo
Anthrax toxin-mediated delivery of a cytotoxic T-cell epitope in vivo
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DOI：
10.1073/pnas.93.22.12531
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发表时间：
1996-10-29
影响因子：
11.1
通讯作者：
Starnbach, MN
中科院分区：
文献类型：
--
作者：
Ballard, JD;Collier, RJ;Starnbach, MN
炭疽毒素的保护性抗原（PA）组分介导毒素的致死因子（LF）和水肿因子进入哺乳动物细胞的胞质区室。LF的氨基末端结构域（LFn; 255个氨基酸）将LF结合至PA，并且当融合至异源蛋白质时，LFn结构域在PA存在下将此类蛋白质递送至细胞质。在本研究中，我们将来自细胞内病原体单核细胞增生李斯特菌的9个氨基酸的细胞毒性T淋巴细胞（CTL）表位（LLO（91-99））融合到LFn，并测量所得LFn-LLO（91-99）融合蛋白在BALB/c小鼠中刺激针对该表位的CTL应答的能力。刺激是PA依赖性的，并且发生与LFn的氨基末端或羧基末端融合的肽。挑战L。在单核细胞增多症中，先前注射LFn-LLO（91-99）和PA的小鼠相对于未免疫的对照小鼠显示脾和肝中的集落形成单位减少。这些结果表明炭疽毒素可用作用于研究和医学应用的CTL-肽递送系统。
The protective antigen (PA) component of anthrax toxin mediates entry of the toxin's lethal factor (LF) and edema factor into the cytosolic compartment of mammalian cells, The amino-terminal domain of LF (LFn; 255 amino acids) binds LF to PA, and when fused to heterologous proteins, the LFn domain delivers such proteins to the cytoplasm in the presence of PA. In the current study, we fused a 9-amino acid cytotoxic T-lymphocyte (CTL) epitope (LLO(91-99)) from an intracellular pathogen, Listeria monocytogenes, to LFn and measured the ability of the resulting LFn-LLO(91-99) fusion protein to stimulate a CTL response against the epitope in BALB/c mice, As little as 300 fmol of fusion could stimulate a response. The stimulation was PA-dependent and occurred with the peptide fused to either the amino terminus or the carboxyl terminus of LFn. Upon challenge with L. monocytogenes, mice previously injected with LFn-LLO(91-99) and PA showed a reduction of colony-forming units in spleen and liver, relative to nonimmunized control mice, These results indicate that anthrax toxin may be useful as a CTL-peptide delivery system for research and medical applications.